STUDIES ON SEED STORAGE PROTEINS OF SOME ECONOMICALLY MINOR PLANTS

Save this PDF as:
 WORD  PNG  TXT  JPG

Size: px
Start display at page:

Download "STUDIES ON SEED STORAGE PROTEINS OF SOME ECONOMICALLY MINOR PLANTS"

Transcription

1 STUDIES ON SEED STORAGE PROTEINS OF SOME ECONOMICALLY MINOR PLANTS THESIS SUBMITTED FOR THE DEGREB OF DOCTOR OF PHILOSOPHY (SCIENCE) OF THE UNIVERSITY OF CALCUTTA 1996 NRISINHA DE, M.Sc DEPARTMENT OF BIOCHEMISTRY BOSE INSTITUTE CALCUTTA

2 CONTENTS Page No INTRODUCTION Seed development Seed grain proteins Characteristic features of seed storage proteins Complexity of seed storage proteins Homologies between storage protein and their genes Storage protein synthesis and deposition Regulation of storage protein synthesis Regulatory sequences of storage protein genes Expression of seed protein genes in transgenic plants Implications of nutritional improvement of seeds by transgenesis Expression of plant genes in prokaryote Importance of economically minor seeds as food sources Aim and Scope of present study 28 MATERIALS AND METHODS Materials Methods RESULTS AND DISCUSSION SECTION -I characterization of 2S albumin from the seeds of Chenopodium album and its accumulation during seed development SECTION -II Characterization of storage proteins from seeds of sterculia foetida SECTION -III Cloning and sequencing of 2S protein gene of Chenopodium album GENERAL DISCUSSION SUMMARY REFERENCES PUBLICATIONS 119 1

3 The work mainly describes the importance of seed storage proteins of two economically minor plants, Chenopodium album and Sterculia foetida. Three nutritionally balanced seed storage proteins were isolated, characterized and their amino acid composition analysed. These balanced proteins may be used as protein food suppliments for animals and human beings. An attempt was made to isolate 2S protein gene of Chenopodium album by conventional method and the gene appears to be a potential candidate for plant improvement by using modern biotechniques. The work is summerised below: t Chapter - I ] 1. Fresh weight of growing Chenopodium album seeds increased linearly from the first week after anthesis upto third week then decreased slowly during dehydration stage. Dry weight of the seeds was found to increase more slowly at the begining and on the subsequent stages it followed a sigmodial pattern. 2. The amount of DNA was almost constant upto second week and then increased slowly until fifth week. The level of RNA increased linearly but slowly upto third week and then rapidly increased to its maximum level at the fourth week after flowering. The accumulation of 99

4 storage proteins followed almost a sigmodial pattern, begining at the second week and continuing upto fifth week. An analysis of SDS-PAGE pattern of seed storage proteins of C^. album at different stages of development revealed that deposition of major classes of seed storage protein started from the second week after flowering. In subsequent days most of the proteins accumulated showed the pattern that of matured processed proteins. A low molecular albumin from Chenopodium album was purified by molecular seive chromatography on Bio-gel P-60 column. It was characterized with respect to its sub-units structure, amino acid compositions and antigenecity. This albumin was found to contain nutritionally balanced amino acid composition comparable to that of recommended values of World Health Organisation (WHO) for an ideal protein. Again this albumin was found to be antigenically homologous with the seed storage proteins of some phylogenetically related species belonging to Chenopodiacae. Amaranthuceae and Basilaceae. but not to that of unrelated dicots. 100

5 [ Chapter II ] Gradual accumulation of major storage proteins in Sterculia foetida seeds were traced by SDS-PAGE analysis. Two proteins, an albumin and a globulin, were purified from S_;_ f oetida seeds through chromatography on sephadex G-150 column and their subunits composition patterns were studied on SDS-PAGE. Their amino acid composition matched closely to an ideal protein milk casein, and was also comparable to the recommended values by World Health Organisation (WHO) for an ideal balanced protein for human consumption. Antibody against the purified globulin was raised in rabbit. Sterculia globulin specific antibody was found to crossreact with seed proteins of phylogenetically related species of Sterculiaceae family, but not with other dicots. [ Chapter - III ] The gene for 2S albumin of Chenopodium album seeds, was chosen as a potential candidate gene for use in seed protein quality improvement by biotechnological approaches, with a view to clone the cdna of this, total RNA was extracted from mid-matured seeds of Chenopodium. Poly A + RNA was purified from extracted 101

6 total RNA by oligo dt-cellulose column chromatography. Poly A + RNA was reversed transcribed to corresponding complementary DNA (cdna) by using reverse transcriptase enzyme (RAV-2), Random hexamer primers were used for cdna synthesis. Complementary DNA (cdna) synthesis was monitored by autoradiography using radiolabled -P 32 datp. A plasmid expression vector pgex-2t was selected for library construction. This plasmid vector can efficiently synthesize forign polypeptide from the inserted gene or gene fragment as fusion protein with Glutathion S Transferase (GST) protein. Recombinant colonies were freshly grown on LB-amp plate overnight were transferred onto a nitrocellulose membrane and induced by 5 mm IPTG. Induced colonies on the membrane were lysed in presence of 5% SDS. The bound bacterial protein on the membrane then probed with antibody and developed according to the Western blot protocol. Two such immunopositive colonies (pgca2s-l and pgca2s-2) were identified by screening the colonies. One colony (pgca2s-l) was further studied in details. The cell lysate from this colony was prepared and run 102

7 on a SDS-PAGE. A fusion protein of around 45 Kd was located on the gel, which was found to crossreact with the antibodies raised against the 2S albumin of Chenopodium by Western blot analysis. Plasmid DNA from this colony was prepared. The insert DNA of 1.0 Kb was separated following EcoR 1/ BamH I digestion. The insert DNA was eluted from low melting agarose gel and cloned in pbsks (+) plasmid vector in corresponding restriction sites (EcoR I and BamH I).The The recombinant pbsks (+) clone was isolated and designated as pbsca2s-l. The sequencing of the DNA insert from the pbsca2s-l plasmid was done by dideoxy chain termination method us ing T 3 and T 7 primers and sequence o f the insert DNA was read. Deduced aminoacid sequence in all possible frames in the forward and reverse complementary sequences were obtained by computer analysis and was compared with published aminoacid sequences of several 2S albumin from different other dicotyledonous plant seeds to arrive at the correct alignment. This cdna clone will be useful in fishing out the desired gene from a Chenopodium genomic library. Further work along this line is in progress. 103

Chapter 8: Recombinant DNA 2002 by W. H. Freeman and Company Chapter 8: Recombinant DNA 2002 by W. H. Freeman and Company

Chapter 8: Recombinant DNA 2002 by W. H. Freeman and Company Chapter 8: Recombinant DNA 2002 by W. H. Freeman and Company Biotechnology and reporter genes Here, a lentivirus is used to carry foreign DNA into chickens. A reporter gene (GFP)indicates that foreign DNA has been successfully transferred. Recombinant DNA continued

More information

Genetics Faculty of Agriculture and Veterinary Medicine

Genetics Faculty of Agriculture and Veterinary Medicine Genetics 10201232 Faculty of Agriculture and Veterinary Medicine Instructor: Dr. Jihad Abdallah Topic 15:Recombinant DNA Technology 1 Recombinant DNA Technology Recombinant DNA Technology is the use of

More information

Molecular Biology Techniques: A Classroom Laboratory Manual THIRD EDITION

Molecular Biology Techniques: A Classroom Laboratory Manual THIRD EDITION Molecular Biology Techniques: A Classroom Laboratory Manual THIRD EDITION Susan Carson Heather B. Miller D.Scott Witherow ELSEVIER AMSTERDAM BOSTON HEIDELBERG LONDON NEW YORK OXFORD PARIS SAN DIEGO SAN

More information

Chapter 12 - DNA Technology

Chapter 12 - DNA Technology Bio 100 DNA Technology 1 Chapter 12 - DNA Technology Among bacteria, there are 3 mechanisms for transferring genes from one cell to another cell: transformation, transduction, and conjugation 1. Transformation

More information

Recombinant DNA Technology

Recombinant DNA Technology PowerPoint Lecture Presentations prepared by Mindy Miller-Kittrell, North Carolina State University C H A P T E R 8 Recombinant DNA Technology The Role of Recombinant DNA Technology in Biotechnology Biotechnology

More information

3. comparison with proteins of known function

3. comparison with proteins of known function Lectures 26 and 27 recombinant DNA technology I. oal of genetics A. historically - easy to isolate total DNA - difficult to isolate individual gene B. recombinant DNA technology C. why get the gene? 1.

More information

The correct answer is c B. Answer b is incorrect. Type II enzymes recognize and cut a specific site, not at random sites.

The correct answer is c B. Answer b is incorrect. Type II enzymes recognize and cut a specific site, not at random sites. 1. A recombinant DNA molecules is one that is a. produced through the process of crossing over that occurs in meiosis b. constructed from DNA from different sources c. constructed from novel combinations

More information

CHAPTER 8 RECOMBINANT DNA and GENETIC ENGINEERING

CHAPTER 8 RECOMBINANT DNA and GENETIC ENGINEERING CHAPTER 8 RECOMBINANT DNA and GENETIC ENGINEERING Questions to be addressed: How are recombinant DNA molecules generated in vitro? How is recombinant DNA amplified? What analytical techniques are used

More information

Common Course Topics Biology 1414: Introduction to Biotechnology I

Common Course Topics Biology 1414: Introduction to Biotechnology I Common Course Topics Biology 1414: Introduction to Biotechnology I Assumptions Students may be enrolled in this course for several reasons; they are enrolled in the Biotechnology Program, they need a science

More information

Chapter 9. Biotechnology and Recombinant DNA Biotechnology and Recombinant DNA

Chapter 9. Biotechnology and Recombinant DNA Biotechnology and Recombinant DNA Chapter 9 Biotechnology and Recombinant DNA Biotechnology and Recombinant DNA Q&A Interferons are species specific, so that interferons to be used in humans must be produced in human cells. Can you think

More information

CHAPTER 14 LECTURE NOTES: RECOMBINANT DNA TECHNOLOGY

CHAPTER 14 LECTURE NOTES: RECOMBINANT DNA TECHNOLOGY CHAPTER 14 LECTURE NOTES: RECOMBINANT DNA TECHNOLOGY I. General Info A. Landmarks in modern genetics 1. Rediscovery of Mendel s work 2. Chromosomal theory of inheritance 3. DNA as the genetic material

More information

Chapter 20: Biotechnology: DNA Technology & Genomics

Chapter 20: Biotechnology: DNA Technology & Genomics Biotechnology Chapter 20: Biotechnology: DNA Technology & Genomics The BIG Questions How can we use our knowledge of DNA to: o Diagnose disease or defect? o Cure disease or defect? o Change/improve organisms?

More information

Recombinant DNA and Biotechnology

Recombinant DNA and Biotechnology Recombinant DNA and Biotechnology Chapter 18 Lecture Objectives What Is Recombinant DNA? How Are New Genes Inserted into Cells? What Sources of DNA Are Used in Cloning? What Other Tools Are Used to Study

More information

Biotechnology and Recombinant DNA

Biotechnology and Recombinant DNA Biotechnology and Recombinant DNA Recombinant DNA procedures - an overview Biotechnology: The use of microorganisms, cells, or cell components to make a product. Foods, antibiotics, vitamins, enzymes Recombinant

More information

Lecture 5 - Bacteriophage Vectors; Lab Practicum 2 (AMG text pp ) September 4, 2001

Lecture 5 - Bacteriophage Vectors; Lab Practicum 2 (AMG text pp ) September 4, 2001 Lecture 5 - Bacteriophage Vectors; Lab Practicum 2 (AMG text pp. 42-55) September 4, 2001 Biology of Bacteriophage Lamda Lambda phage is a bacterial virus that infects E. coli, and depending on early events

More information

Lecture 13. Molecular Cloning

Lecture 13. Molecular Cloning Lecture 13 Molecular Cloning Recombinant DNA technology depends on the ability to produce large numbers of identical DNA molecules (clones). Clones are typically generated by placing a DNA fragment of

More information

Biotechnology and Recombinant DNA (Chapter 9) Lecture Materials for Amy Warenda Czura, Ph.D. Suffolk County Community College

Biotechnology and Recombinant DNA (Chapter 9) Lecture Materials for Amy Warenda Czura, Ph.D. Suffolk County Community College Biotechnology and Recombinant DNA (Chapter 9) Lecture Materials for Amy Warenda Czura, Ph.D. Suffolk County Community College Primary Source for figures and content: Eastern Campus Tortora, G.J. Microbiology

More information

Recombinant DNA technology (genetic engineering) involves combining genes from different sources into new cells that can express the genes.

Recombinant DNA technology (genetic engineering) involves combining genes from different sources into new cells that can express the genes. Recombinant DNA technology (genetic engineering) involves combining genes from different sources into new cells that can express the genes. Recombinant DNA technology has had-and will havemany important

More information

Chapter 20: Biotechnology

Chapter 20: Biotechnology Name Period The AP Biology exam has reached into this chapter for essay questions on a regular basis over the past 15 years. Student responses show that biotechnology is a difficult topic. This chapter

More information

restriction enzymes 350 Home R. Ward: Spring 2001

restriction enzymes 350 Home R. Ward: Spring 2001 restriction enzymes 350 Home Restriction Enzymes (endonucleases): molecular scissors that cut DNA Properties of widely used Type II restriction enzymes: recognize a single sequence of bases in dsdna, usually

More information

Recombinant DNA & Genetic Engineering. Tools for Genetic Manipulation

Recombinant DNA & Genetic Engineering. Tools for Genetic Manipulation Recombinant DNA & Genetic Engineering g Genetic Manipulation: Tools Kathleen Hill Associate Professor Department of Biology The University of Western Ontario Tools for Genetic Manipulation DNA, RNA, cdna

More information

Integrated Protein Services

Integrated Protein Services Integrated Protein Services Custom protein expression & purification Version DC04-0012 Expression strategy The first step in the recombinant protein generation process is to design an appropriate expression

More information

Chapter 10 Manipulating Genes

Chapter 10 Manipulating Genes How DNA Molecules Are Analyzed Chapter 10 Manipulating Genes Until the development of recombinant DNA techniques, crucial clues for understanding how cell works remained lock in the genome. Important advances

More information

Expression and Purification of Recombinant Protein in bacteria and Yeast. Presented By: Puspa pandey, Mohit sachdeva & Ming yu

Expression and Purification of Recombinant Protein in bacteria and Yeast. Presented By: Puspa pandey, Mohit sachdeva & Ming yu Expression and Purification of Recombinant Protein in bacteria and Yeast Presented By: Puspa pandey, Mohit sachdeva & Ming yu DNA Vectors Molecular carriers which carry fragments of DNA into host cell.

More information

DNA TECHNOLOGY- methods for studying and manipulating genetic material.

DNA TECHNOLOGY- methods for studying and manipulating genetic material. 1 DNA TECHNOLOGY- methods for studying and manipulating genetic material. BIOTECHNOLOGY, the manipulation of organisms or their components to make useful products. Biotechnology today usually refers to

More information

Recombinant DNA Technology

Recombinant DNA Technology Recombinant DNA Technology Dates in the Development of Gene Cloning: 1965 - plasmids 1967 - ligase 1970 - restriction endonucleases 1972 - first experiments in gene splicing 1974 - worldwide moratorium

More information

TECH NOTE Characterization of CRISPR/Cas9 introduced Mutations using the Guide it Indel Identification Kit

TECH NOTE Characterization of CRISPR/Cas9 introduced Mutations using the Guide it Indel Identification Kit TECH NOTE Characterization of CRISPR/Cas9 introduced Mutations using the Guide it Indel Identification Kit Streamlined method for characterizing the variety of indels introduced by genome editing technologies:

More information

Integrated Protein Services

Integrated Protein Services Integrated Protein Services Custom protein expression & purification Last date of revision June 2015 Version DC04-0013 www.iba-lifesciences.com Expression strategy The first step in the recombinant protein

More information

Antibody Production Price List

Antibody Production Price List Antibody Production Price List Presenting Insight Biotechnology s price list for custom polyclonal and monoclonal antibody production services. We are happy to tailor individual packages towards the specific

More information

Biotechnology: DNA Technology & Genomics

Biotechnology: DNA Technology & Genomics Chapter 20. Biotechnology: DNA Technology & Genomics 2003-2004 The BIG Questions How can we use our knowledge of DNA to: diagnose disease or defect? cure disease or defect? change/improve organisms? What

More information

Lecture 36: Basics of DNA Cloning-II

Lecture 36: Basics of DNA Cloning-II Lecture 36: Basics of DNA Cloning-II Note: Before starting this lecture students should have completed Lecture 35 Sequential steps involved in DNA cloning using plasmid DNA as vector: Molecular cloning

More information

Tools and Techniques. Chapter 10. Genetic Engineering. Restriction endonuclease. 1. Enzymes

Tools and Techniques. Chapter 10. Genetic Engineering. Restriction endonuclease. 1. Enzymes Chapter 10. Genetic Engineering Tools and Techniques 1. Enzymes 2. 3. Nucleic acid hybridization 4. Synthesizing DNA 5. Polymerase Chain Reaction 1 2 1. Enzymes Restriction endonuclease Ligase Reverse

More information

Plasmid Isolation. Prepared by Latifa Aljebali Office: Building 5, 3 rd floor, 5T250

Plasmid Isolation. Prepared by Latifa Aljebali Office: Building 5, 3 rd floor, 5T250 Plasmid Isolation Prepared by Latifa Aljebali Office: Building 5, 3 rd floor, 5T250 Plasmid Plasmids are small, double strand, closed circular DNA molecules. Isolated from bacterial cells. Replicate independently

More information

Recipient Cell. DNA Foreign DNA. Recombinant DNA

Recipient Cell. DNA Foreign DNA. Recombinant DNA Module 4B Biotechnology In this module, we will examine some of the techniques scientists have developed to study and manipulate the DNA of living organisms. Objective # 7 Explain what genetic recombination

More information

HiPer RT-PCR Teaching Kit

HiPer RT-PCR Teaching Kit HiPer RT-PCR Teaching Kit Product Code: HTBM024 Number of experiments that can be performed: 5 Duration of Experiment: Protocol: 4 hours Agarose Gel Electrophoresis: 45 minutes Storage Instructions: The

More information

Recombinant DNA Unit Exam

Recombinant DNA Unit Exam Recombinant DNA Unit Exam Question 1 Restriction enzymes are extensively used in molecular biology. Below are the recognition sites of two of these enzymes, BamHI and BclI. a) BamHI, cleaves after the

More information

Just the Facts: A Basic Introduction to the Science Underlying NCBI Resources

Just the Facts: A Basic Introduction to the Science Underlying NCBI Resources 1 of 8 11/7/2004 11:00 AM National Center for Biotechnology Information About NCBI NCBI at a Glance A Science Primer Human Genome Resources Model Organisms Guide Outreach and Education Databases and Tools

More information

pcas-guide System Validation in Genome Editing

pcas-guide System Validation in Genome Editing pcas-guide System Validation in Genome Editing Tagging HSP60 with HA tag genome editing The latest tool in genome editing CRISPR/Cas9 allows for specific genome disruption and replacement in a flexible

More information

Today-applications: Medicine-better health Pharmaceutical-production of antibiotics Foods-wine, cheese, beer Agriculture-selective breeding

Today-applications: Medicine-better health Pharmaceutical-production of antibiotics Foods-wine, cheese, beer Agriculture-selective breeding I. Genetic Engineering modification of DNA of organisms to produce new genes with new characteristics -genes are small compared to chromosomes -need methods to get gene-sized pieces of DNA -direct manipulation

More information

First Strand cdna Synthesis

First Strand cdna Synthesis 380PR 01 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name First Strand cdna Synthesis (Cat. # 786 812) think proteins! think G-Biosciences

More information

05_02_DNA.jpg. covalent bonds H bonds. polarity; antiparallel

05_02_DNA.jpg. covalent bonds H bonds. polarity; antiparallel Animal Cell 05_02_DNA.jpg covalent bonds H bonds polarity; antiparallel 05_06_compl_pairs.jpg H bonds between a purine and a pyrimidine fits the space; complementary bases; which more stable? 05_07_base

More information

Lecture 10. mrna: Transcription Translation Start Translation Stop Transcription Start (AUG) (UAG, UAA, or UGA) Terminator S-D Sequence

Lecture 10. mrna: Transcription Translation Start Translation Stop Transcription Start (AUG) (UAG, UAA, or UGA) Terminator S-D Sequence Lecture 10 Analysis of Gene Sequences Anatomy of a bacterial gene: Promoter Coding Sequence (no stop codons) mrna: Transcription Translation Start Translation Stop Transcription Start (AUG) (UAG, UAA,

More information

Protein immunoblotting

Protein immunoblotting Protein immunoblotting (Western blotting) Dr. Serageldeen A. A. Sultan Lecturer of virology Dept. of Microbiology SVU, Qena, Egypt seaas@lycos.com Western blotting -It is an analytical technique used to

More information

RECOMBINANT DNA TECHNOLOGY

RECOMBINANT DNA TECHNOLOGY RECOMBINANT DNA TECHNOLOGY By; Dr. Adeel Chaudhary 2 nd yr Molecular Genetics Medical Technology College of Applied Medical Sciences Recombinant DNA is a form of artificial DNA that is made through the

More information

How many of you have checked out the web site on protein-dna interactions?

How many of you have checked out the web site on protein-dna interactions? How many of you have checked out the web site on protein-dna interactions? Example of an approximately 40,000 probe spotted oligo microarray with enlarged inset to show detail. Find and be ready to discuss

More information

CAP BIOINFORMATICS Su-Shing Chen CISE. 10/5/2005 Su-Shing Chen, CISE 1

CAP BIOINFORMATICS Su-Shing Chen CISE. 10/5/2005 Su-Shing Chen, CISE 1 CAP 5510-8 BIOINFORMATICS Su-Shing Chen CISE 10/5/2005 Su-Shing Chen, CISE 1 Genomic Mapping & Mapping Databases High resolution, genome-wide maps of DNA markers. Integrated maps, genome catalogs and comprehensive

More information

Biotechnology. Biotechnology s Laboratories. Lab Name Location Person in Charge Programs Served Courses Served. Biotechnology Department

Biotechnology. Biotechnology s Laboratories. Lab Name Location Person in Charge Programs Served Courses Served. Biotechnology Department Biotechnology s oratories Biotechnology Name Location Person in Charge Programs Served Courses Served General Biology W12-039 Zahra Yassin General Microbiology M12-132 Aisha Echtibi General (Basic Course)

More information

Pharmaceutical Biotechnology. Recombinant DNA technology Western blotting and SDS-PAGE

Pharmaceutical Biotechnology. Recombinant DNA technology Western blotting and SDS-PAGE Pharmaceutical Biotechnology Recombinant DNA technology Western blotting and SDS-PAGE Recombinant DNA Technology Protein Synthesis Western Blot Western blots allow investigators to determine the molecular

More information

Name Class Date WHAT I KNOW. organisms with specific traits for certain functions. For example, some plants provide food.

Name Class Date WHAT I KNOW. organisms with specific traits for certain functions. For example, some plants provide food. Genetic Engineering Science as a Way of Knowing Q: How and why do scientists manipulate DNA in living cells? 15.1 How do humans take advantage of naturally occurring variation among organisms? WHAT I KNOW

More information

QuickClean 96-Well Plasmid Miniprep Kit

QuickClean 96-Well Plasmid Miniprep Kit QuickClean 96-Well Plasmid Miniprep Kit L00237 Technical Manual No. TM 0231 Version 0712007 I Description.. 1 II Kit Contents.. 1 III Applications 2 IV Key Features.. 2 V Storage.. 2 VI Plasmid Miniprep

More information

Solutions to Problem Set 5

Solutions to Problem Set 5 MIT Biology Department 7.012: Introductory Biology - Fall 2004 Instructors: Professor Eric Lander, Professor Robert A. Weinberg, Dr. Claudette Gardel Question 1 Solutions to 7.012 Problem Set 5 Restriction

More information

Solutions for Recombinant DNA Unit Exam

Solutions for Recombinant DNA Unit Exam Solutions for Recombinant DNA Unit Exam Question 1 Restriction enzymes are extensively used in molecular biology. Below are the recognition sites of two of these enzymes, BamHI and BclI. a) BamHI, cleaves

More information

Biochem 717 Gene Cloning. Prof Amer Jamil Dept of Biochemistry University of Agriculture Faisalabad

Biochem 717 Gene Cloning. Prof Amer Jamil Dept of Biochemistry University of Agriculture Faisalabad Biochem 717 Gene Cloning Prof Amer Jamil Dept of Biochemistry University of Agriculture Faisalabad How to construct a recombinant DNA molecule? DNA isolation Cutting of DNA molecule with the help of restriction

More information

Chapter 2 Biological Inventions

Chapter 2 Biological Inventions Note: When any ambiguity of interpretation is found in this provisional translation, the Japanese text shall prevail. Chapter 2 Biological Inventions (Applied to any patent applications filed on or after

More information

Aviva Systems Biology

Aviva Systems Biology Aviva Custom Antibody Service and Price Mouse Monoclonal Antibody Service Package Number Description Package Contents Time Price Customer provides antigen protein $6,174 Monoclonal package1 (From protein

More information

pglo Bacterial Transformation

pglo Bacterial Transformation Introduction pglo Bacterial Transformation Biotechnology refers to technology used to manipulate DNA. The procedures are often referred to as genetic engineering. DNA is the genetic material of all living

More information

Department of Biology Sample

Department of Biology Sample Syllabus BIOTECHNOLOGY Spring 2013 Instructor: Atanu Duttaroy, Professor Tel: 202-806-5362 Email: aduttaroy@howard.edu Office: Room 336, Just Hall Teaching Assistant: Mr. Subhas Mukherjee Lecture: Room

More information

Overview of the Recombinant DNA technology- the process of subcloning a foreign gene into the plasmid vector puc19

Overview of the Recombinant DNA technology- the process of subcloning a foreign gene into the plasmid vector puc19 Health and Life Sciences Faculty Course Title: Biological and Forensic Science Module code: 216 BMS Module Title: Molecular Genetics Overview of the Recombinant DNA technology- the process of subcloning

More information

HCS604.03 Exercise 1 Dr. Jones Spring 2005. Recombinant DNA (Molecular Cloning) exercise:

HCS604.03 Exercise 1 Dr. Jones Spring 2005. Recombinant DNA (Molecular Cloning) exercise: HCS604.03 Exercise 1 Dr. Jones Spring 2005 Recombinant DNA (Molecular Cloning) exercise: The purpose of this exercise is to learn techniques used to create recombinant DNA or clone genes. You will clone

More information

AP BIOLOGY 2009 SCORING GUIDELINES (Form B)

AP BIOLOGY 2009 SCORING GUIDELINES (Form B) AP BIOLOGY 2009 SCORING GUIDELINES (Form B) Question 1 Describe how a plasmid can be genetically modified to include a piece of foreign DNA that alters the phenotype of bacterial cells transformed with

More information

CCR Biology - Chapter 9 Practice Test - Summer 2012

CCR Biology - Chapter 9 Practice Test - Summer 2012 Name: Class: Date: CCR Biology - Chapter 9 Practice Test - Summer 2012 Multiple Choice Identify the choice that best completes the statement or answers the question. 1. Genetic engineering is possible

More information

KMS-Specialist & Customized Biosimilar Service

KMS-Specialist & Customized Biosimilar Service KMS-Specialist & Customized Biosimilar Service 1. Polyclonal Antibody Development Service KMS offering a variety of Polyclonal Antibody Services to fit your research and production needs. we develop polyclonal

More information

Aviva Systems Biology

Aviva Systems Biology Aviva Custom Antibody Services and Prices Rabbit Polyclonal Antibody Service Package Number Description Package Contents Time Price Polyclonal package 1 (From protein to antiserum) Polyclonal package 2

More information

STANDARD 2 Students will demonstrate appropriate safety procedures and equipment use in the laboratory.

STANDARD 2 Students will demonstrate appropriate safety procedures and equipment use in the laboratory. BIOTECHNOLOGY Levels: 11-12 Units of Credit: 1.0 CIP Code: 51.1201 Prerequisite: Biology or Chemistry Skill Certificates: #708 COURSE DESCRIPTION is an exploratory course designed to create an awareness

More information

CUSTOM ANTIBODIES. Fully customised services: rat and murine monoclonals, rat and rabbit polyclonals, antibody characterisation, antigen preparation

CUSTOM ANTIBODIES. Fully customised services: rat and murine monoclonals, rat and rabbit polyclonals, antibody characterisation, antigen preparation CUSTOM ANTIBODIES Highly competitive pricing without compromising quality. Rat monoclonal antibodies for the study of gene expression and proteomics in mice and in mouse models of human diseases available.

More information

The Cloning and Expression of Rat Insulin2like Growth Factor Gene

The Cloning and Expression of Rat Insulin2like Growth Factor Gene ,29 (12) :1063 1067,2002 Acta Genetica Sinica,,, (, 300070) : DNA,PCR ( IGF2 ) 2 3, 2 3 3 5 40, PCR, 210bp IGF2 puc18 pgex2igf2, DH5 SDS2PAGE Western blot, : ; ; ; :Q784 :A :0379-4172 (2002) 12-1063 -

More information

A polynucleotide consisting of the nucleotide sequence of SEQ ID NO:2.

A polynucleotide consisting of the nucleotide sequence of SEQ ID NO:2. Case A A polynucleotide consisting of the nucleotide sequence of SEQ ID NO:1 nucleotide sequence of SEQ ID NO:1 is one of the sequences which were analyzed using an automated DNA sequencer The sequences

More information

'LVFXVVLRQ $UUD\HGF'1$H[SUHVVLRQOLEUDULHV 5RERWWHFKQRORJ\DQGDUUD\HGOLEUDULHV

'LVFXVVLRQ $UUD\HGF'1$H[SUHVVLRQOLEUDULHV 5RERWWHFKQRORJ\DQGDUUD\HGOLEUDULHV Discussion 74 'LVFXVVLRQ This study describes arrayed cdna libraries as a source of clonally expressed recombinant proteins which can be directly linked to clones characterised and identified by DNA hybridisation

More information

Creation of a Bacterial Cell Controlled by a Chemically Synthesized Genome Gibson et al. (2010)

Creation of a Bacterial Cell Controlled by a Chemically Synthesized Genome Gibson et al. (2010) Creation of a Bacterial Cell Controlled by a Chemically Synthesized Genome Gibson et al. (2010) In 1977 Sanger and his colleagues were the first who sequenced the complete DNA genome of a phage. From 1977

More information

Nucleic Acid Techniques in Bacterial Systematics

Nucleic Acid Techniques in Bacterial Systematics Nucleic Acid Techniques in Bacterial Systematics Edited by Erko Stackebrandt Department of Microbiology University of Queensland St Lucia, Australia and Michael Goodfellow Department of Microbiology University

More information

Chapter 3 Contd. Western blotting & SDS PAGE

Chapter 3 Contd. Western blotting & SDS PAGE Chapter 3 Contd. Western blotting & SDS PAGE Western Blot Western blots allow investigators to determine the molecular weight of a protein and to measure relative amounts of the protein present in different

More information

Microbiology / Active Lecture Questions Chapter 9 Biotechnology & Recombinant DNA 1 Chapter 9 Biotechnology & Recombinant DNA

Microbiology / Active Lecture Questions Chapter 9 Biotechnology & Recombinant DNA 1 Chapter 9 Biotechnology & Recombinant DNA 1 2 Restriction enzymes were first discovered with the observation that a. DNA is restricted to the nucleus. b. phage DNA is destroyed in a host cell. c. foreign DNA is kept out of a cell. d. foreign DNA

More information

Genetic transformation literally means change caused by genes.

Genetic transformation literally means change caused by genes. pglo Bacterial Transformation Practical What is transformation? Genetic transformation literally means change caused by genes. It occurs when a cell takes up (takes inside) and expresses a new piece of

More information

Lecture 13: DNA Technology. DNA Sequencing. DNA Sequencing Genetic Markers - RFLPs polymerase chain reaction (PCR) products of biotechnology

Lecture 13: DNA Technology. DNA Sequencing. DNA Sequencing Genetic Markers - RFLPs polymerase chain reaction (PCR) products of biotechnology Lecture 13: DNA Technology DNA Sequencing Genetic Markers - RFLPs polymerase chain reaction (PCR) products of biotechnology DNA Sequencing determine order of nucleotides in a strand of DNA > bases = A,

More information

MOLECULAR GENETICS GENETIC ENGINEERING RECOMBINANT DNA. Molecular Genetics Activity #6 page 1

MOLECULAR GENETICS GENETIC ENGINEERING RECOMBINANT DNA. Molecular Genetics Activity #6 page 1 AP BIOLOGY MOLECULAR GENETICS ACTIVITY #6 NAME DATE HOUR RECOMBINANT DNA GENETIC ENGINEERING Molecular Genetics Activity #6 page 1 GENETIC ENGINEERING Molecular Genetics Activity #6 page 2 PART I: PRODUCING

More information

Common Course Topics Biology 1406: Cell and Molecular Biology

Common Course Topics Biology 1406: Cell and Molecular Biology Common Course Topics Biology 1406: Cell and Molecular Biology 1. Introduction to biology --the scientific study of organisms --properties of life --assumptions, methods and limitations of science --underlying

More information

Name Class Date. KEY CONCEPT Mutations are changes in DNA that may or may not affect phenotype. frameshift mutation

Name Class Date. KEY CONCEPT Mutations are changes in DNA that may or may not affect phenotype. frameshift mutation Unit 7 Study Guide Section 8.7: Mutations KEY CONCEPT Mutations are changes in DNA that may or may not affect phenotype. VOCABULARY mutation point mutation frameshift mutation mutagen MAIN IDEA: Some mutations

More information

Bacterial Transformation and Plasmid Purification. Chapter 5: Background

Bacterial Transformation and Plasmid Purification. Chapter 5: Background Bacterial Transformation and Plasmid Purification Chapter 5: Background History of Transformation and Plasmids Bacterial methods of DNA transfer Transformation: when bacteria take up DNA from their environment

More information

Many cells will not take up plasmid during transformation Cells with plasmid can be identified because original plasmid contained gene for antibiotic

Many cells will not take up plasmid during transformation Cells with plasmid can be identified because original plasmid contained gene for antibiotic Many cells will not take up plasmid during transformation Cells with plasmid can be identified because original plasmid contained gene for antibiotic resistance (ampicillin) Use medium with ampicillin

More information

DNA Fingerprinting. Unless they are identical twins, individuals have unique DNA

DNA Fingerprinting. Unless they are identical twins, individuals have unique DNA DNA Fingerprinting Unless they are identical twins, individuals have unique DNA DNA fingerprinting The name used for the unambiguous identifying technique that takes advantage of differences in DNA sequence

More information

Microbiology Laboratory: MOLECULAR IDENTIFICATION OF UNKNOWN BACTERIA

Microbiology Laboratory: MOLECULAR IDENTIFICATION OF UNKNOWN BACTERIA Microbiology Laboratory: MOLECULAR IDENTIFICATION OF UNKNOWN BACTERIA Classical Microbiology courses are typically structured to introduce the identification of bacterial species using a series of biochemical

More information

Biotechnology. Selective breeding Use of microbes (bacteria & yeast)

Biotechnology. Selective breeding Use of microbes (bacteria & yeast) Biotechnology bio and technology The use of living organisms to solve problems or make useful products. Biotechnology has been practiced for the last 10,000 years. Selective breeding Use of microbes (bacteria

More information

RNA-related Products

RNA-related Products RNA-related Products TranscriptME RNA kit: Ideal choice for obtaining high yields of full-length cdna for RT-qPCR assays Suitable for as low RNA amount as 10 pg p p Convenient, reliable and cost-effective

More information

Protein Expression. A Practical Approach J. HIGGIN S

Protein Expression. A Practical Approach J. HIGGIN S Protein Expression A Practical Approach S. J. HIGGIN S B. D. HAMES List of contributors Abbreviations xv Xvi i 1. Protein expression in mammalian cell s Marlies Otter-Nilsson and Tommy Nilsso n 1. Introduction

More information

I. Gene transfer and genetic engineering (Chapter 8) A. General concepts 1. Genetic information is contained in the nucleotide sequence of DNA

I. Gene transfer and genetic engineering (Chapter 8) A. General concepts 1. Genetic information is contained in the nucleotide sequence of DNA I. Gene transfer and genetic engineering (Chapter 8) A. General concepts 1. Genetic information is contained in the nucleotide sequence of DNA (sometimes RNA). Amino acids are specified by a triplet codon.

More information

2. Enzymes that cleave DNA at specific sites are called.

2. Enzymes that cleave DNA at specific sites are called. Biotechnology 1. The most recent techniques developed in the biological sciences allow the manipulation of DNA with the ultimate goal of intervening directly with the fate of organisms. 2. Enzymes that

More information

RNA-related Products

RNA-related Products RNA-related Products TRANSCRIPTME RNA kit: Ideal choice for obtaining high yields of full-length cdna for RT-qPCR assays Suitable for as low RNA amount as 10 pg p p Convenient, reliable and cost-effective

More information

INTERNATIONAL CONFERENCE ON HARMONISATION OF TECHNICAL REQUIREMENTS FOR REGISTRATION OF PHARMACEUTICALS FOR HUMAN USE Q5B

INTERNATIONAL CONFERENCE ON HARMONISATION OF TECHNICAL REQUIREMENTS FOR REGISTRATION OF PHARMACEUTICALS FOR HUMAN USE Q5B INTERNATIONAL CONFERENCE ON HARMONISATION OF TECHNICAL REQUIREMENTS FOR REGISTRATION OF PHARMACEUTICALS FOR HUMAN USE ICH HARMONISED TRIPARTITE GUIDELINE QUALITY OF BIOTECHNOLOGICAL PRODUCTS: ANALYSIS

More information

Genetic Engineering and Biotechnology

Genetic Engineering and Biotechnology 1 So, what is biotechnology?? The use of living organisms to carry out defined chemical processes for industrial or commercial application. The office of Technology Assessment of the U.S. Congress defines

More information

11/19/2008. Gene analysis. Sequencing PCR. Northern-blot RT PCR. Western-blot Sequencing. in situ hybridization. Southern-blot

11/19/2008. Gene analysis. Sequencing PCR. Northern-blot RT PCR. Western-blot Sequencing. in situ hybridization. Southern-blot Recombinant technology Gene analysis Sequencing PCR RNA Northern-blot RT PCR Protein Western-blot Sequencing Southern-blot in situ hybridization in situ hybridization Function analysis Histochemical analysis

More information

How Do You Make A Transgenic Plant?

How Do You Make A Transgenic Plant? Home Page News Updates History of Plant Breeding What Are Transgenic Plants? How Do You Make A Transgenic Plant? Introduction to DNA Locating genes for plant traits Designing genes for insertion Transformation

More information

BSCI410-Liu/SP07 Exam #2 Apr. 5, 2007

BSCI410-Liu/SP07 Exam #2 Apr. 5, 2007 Your Name: KEY UID# 1. (20 points) Dr. Liu has isolated a recessive Arabidopsis mutation; mutants homozygous for this mutation produce small seeds. She named this mutant tiny. To map and clone the corresponding

More information

50 g 650 L. *Average yields will vary depending upon a number of factors including type of phage, growth conditions used and developmental stage.

50 g 650 L. *Average yields will vary depending upon a number of factors including type of phage, growth conditions used and developmental stage. 3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Phage DNA Isolation Kit Product # 46800, 46850 Product Insert

More information

GENE CLONING AND RECOMBINANT DNA TECHNOLOGY

GENE CLONING AND RECOMBINANT DNA TECHNOLOGY GENE CLONING AND RECOMBINANT DNA TECHNOLOGY What is recombinant DNA? DNA from 2 different sources (often from 2 different species) are combined together in vitro. Recombinant DNA forms the basis of cloning.

More information

BIOLOGICAL BACKGROUND THE CENTRAL DOGMA OF MOLECULAR BIOLOGY

BIOLOGICAL BACKGROUND THE CENTRAL DOGMA OF MOLECULAR BIOLOGY BIOLOGICAL BACKGROUND Central Dogma DNA and RNA Structure Replication, Transcription and Translation Techniques of Molecular Genetics Using restriction enzymes Using PCR THE CENTRAL DOGMA OF MOLECULAR

More information

Protein Purification and Analysis

Protein Purification and Analysis Protein Purification and Analysis Numbers of genes: Humans ~40,000 genes Yeast ~6000 genes Bacteria ~3000 genes Solubility of proteins important for purification: 60-80% soluble, 20-40% membrane Some proteins

More information

SUMOstar Gene Fusion Technology for Secretory Expression in Mammalian Cells

SUMOstar Gene Fusion Technology for Secretory Expression in Mammalian Cells SUMOstar Gene Fusion Technology for Secretory Expression in Mammalian Cells NEW METHODS FOR ENHANCING FUNCTIONAL PROTEIN EXPRESSION AND PURIFICATION IN MAMMALIAN CELLS White Paper June 2007 LifeSensors

More information

MOLECULAR BIOLOGY OVERVIEW NUCLEIC ACIDS: THE BASICS

MOLECULAR BIOLOGY OVERVIEW NUCLEIC ACIDS: THE BASICS MOLECULAR BIOLOGY OVERVIEW NUCLEIC ACIDS: THE BASICS Richard L. Hodinka, Ph.D. University of South Carolina School of Medicine Greenville Greenville Health System, Greenville, SC hodinka@greenvillemed.sc.edu

More information

CompleteⅡ 1st strand cdna Synthesis Kit

CompleteⅡ 1st strand cdna Synthesis Kit Instruction Manual CompleteⅡ 1st strand cdna Synthesis Kit Catalog # GM30401, GM30402 Green Mountain Biosystems. LLC Web: www.greenmountainbio.com Tel: 800-942-1160 Sales: Sales@ greenmountainbio.com Support:

More information

Molecular Cloning, Product Brochure

Molecular Cloning, Product Brochure , Product Brochure Interest in any of the products, request or order them at Bio-Connect. Bio-Connect B.V. T NL +31 (0)26 326 44 50 T BE +32 (0)2 503 03 48 Begonialaan 3a F NL +31 (0)26 326 44 51 F BE

More information